We have discovered a new spontaneous circadian mutation in Syrian hamsters, which confers a robust phenotype on the animals'locomotor rhythmicity. The free running period of animals homozygous for the mutation is approximately 28 hours, 4 hours per daily cycle longer than that of wild type animals. Such mutations, most produced in mice by chemical mutagenesis, have been critical in working out what we know thus far about the molecular mechanism that generates cell autonomous circadian rhythmicity in mammals, but our knowledge is far from complete and it is clear that additional unidentified genes must contribute to its function. Furthermore, because hamsters provide many advantages over mice for behavioral and physiological studies, this new mutation provides a unique opportunity for studies of the circadian clock mechanism in mammals. We propose to analyze this new mutation at 3 levels: genetic, behavioral and molecular. Genetic analysis will test our assumption, based on preliminary data, that the mutation is a single, autosomal co-dominant allele. The work in the Menaker laboratory will be focused on the behavioral phenotype and will explore the responses of wild type, heterozygous and homozygous littermates of both sexes to constant darkness, constant light, single light pulses, photoperiods with different light/dark ratis and entraining cycles with different periods, These data will address the role of the mutant gene in the circadian molecular mechanism as well as its impact on the organism's physiology. The Green lab will perform molecular and biochemical analysis of these mutants to determine whether this mutation affects the core intracellular oscillator mechanism or some system-level aspect, such as intercellular coupling. The nature of the mutation will be addressed by sequencing of the known circadian genes, and by analysis of their expression levels. This will also lay important groundwork for the eventual identification of the mutation by whole-genome sequencing. Circadian rhythms modulate much of normal physiology and behavior and circadian disruptions increase vulnerability to a variety of environmental insults. Knowledge of the underlying mechanism is essential to controlling these deleterious effects.

Public Health Relevance

The network of circadian oscillators that stretches from the brain to every structure in periphery has evolved to maintain internal temporal order in the natural environment. However, this system is stressed and internal temporal order is disrupted by many of the changes that humans have made to their own unnatural environments. In order to mitigate the deleterious effects of circadian disruption on human health, it is necessary to understand the intact and the disrupted system in molecular detail.